Immunofluorescence Analysis of FIP200 in NPCs

Immunofluorescence analysis was performed as previously described^{37 (link)}. First, 4% paraformaldehyde was used to fix NPCs, and then 0.5% Triton X-100 in PBS was used to permeabilize. The slides were washed in PBS and blocked with 2% bovine serum albumin (BSA) in PBS for 2 h at 37 °C, and then incubated with anti-FIP200 (1:50) (Proteintech) for 10 h. After washing twice, the slides were then incubated with goat anti-rabbit antibody (CST) at 37 °C for 1 h. Nuclei were then co-stained with 0.1 g/ml DAPI (Beyotime, Nantong, China), and images were captured under a microscope (Olympus, BX53; Melville, NY, USA).